Structure-activity relationship study of mesyl and busyl phosphoramidate antisense oligonucleotides for unaided and PSMA-mediated uptake into prostate cancer cells

Phosphorothioate (PS) group is a key component of a majority of FDA approved oligonucleotide drugs that increase stability to nucleases whilst maintaining interactions with many proteins, including RNase H in the case of antisense oligonucleotides (ASOs). At the same time, uniform PS modification increases nonspecific protein binding that can trigger toxicity and pro-inflammatory effects, so discovery and characterization of alternative phosphate mimics for RNA therapeutics is an actual task. Here we evaluated the effects of the introduction of several N-alkane sulfonyl phosphoramidate groups such as mesyl (methanesulfonyl) or busyl (1-butanesulfonyl) phosphoramidates into gapmer ASOs on the efficiency and pattern of RNase H cleavage, cellular uptake in vitro, and intracellular localization. Using Malat1 lncRNA as a target, we have identified patterns of mesyl or busyl modifications in the ASOs for optimal knockdown in vitro. Combination of the PSMA ligand-mediated delivery with optimized mesyl and busyl ASOs resulted in the efficient target depletion in the prostate cancer cells. Our study demonstrated that other N-alkanesulfonyl phosphoramidate groups apart from a known mesyl phosphoramidate can serve as an essential component of mixed backbone gapmer ASOs to reduce drawbacks of uniformly PS-modified gapmers, and deserve further investigation in RNA therapeutics.

Авторы
Sergeeva O.1 , Akhmetova E.1 , Dukova S.1 , Beloglazkina E.2 , Uspenskaya A.2 , Machulkin A. 2, 3 , Stetsenko D.4 , Zatsepin T.2
Журнал
Издательство
Frontiers Media S. A
Язык
Английский
Страницы
1342178
Статус
Опубликовано
Том
12
Год
2024
Организации
  • 1 Skolkovo Institute of Science and Technology, Moscow
  • 2 Department of Chemistry, Lomonosov Moscow State University, Moscow
  • 3 Department for Biochemistry, People's Friendship University of Russia Named after Patrice Lumumba (RUDN University), Moscow
  • 4 Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk
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